The present disclosure relates generally to methods for cell transfection and regulating cellular behavior. More particularly, the present disclosure relates to methods of using biodegradable mineral coatings for non-viral cell transfection and regulating cellular behavior.
Transfection is the process of introducing nucleic acids into cells. Various transfection strategies are available that generally involve opening transient pores in the cell membrane to allow the uptake of material by the cell. Two broad categories of transfection include chemical-based transfection methods and non-chemical transfection methods. Chemical-based transfection methods include, for example, calcium phosphate-based transfection and liposome-based transfection. In calcium phosphate-based transfection, a buffer containing phosphate ions is combined with a calcium chloride solution containing the DNA to be transfected to form calcium-phosphate precipitates that bind DNA. A suspension of the precipitate is added to cells, which take up the precipitate and DNA. In liposome-based transfection, DNA is incorporated into liposomes that fuse with the cell membrane to release the DNA into the cells.
The deposition of calcium phosphate (CaP)-based materials on various substrates has been used to develop bioactive interfaces for studying its interaction with bone-forming cells. In addition, CaP mineral substrates bind and release biological molecules (e.g., DNA). Studies have also been conducted to investigate the role of CaP mineral properties (physical and chemical) in regulating cellular behavior, including stem cell attachment, proliferation, and differentiation. However, these previous studies have not been capable of systematically studying CaP mineral effects on stem cell proliferation and differentiation. Taken together, the potential impact of these materials on non-viral transfection and stem cell behavior as well as the inherent complexity of CaP requires novel strategies to identify useful CaP mineral coatings. Accordingly, there exists a need to develop non-viral transfection methods and systems for use with these methods.